User:DavidC/16 September 2009

From 2009.igem.org

framless



August
MTWTFSS
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
October
MTWTFSS
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31


Contents

Wednesday the 16th

DNA extraction

DNA extraction from bio-brick R0040+B0030 and P1002+C0040 from 500mL of E.coli DH5alpha growth in LB medium.

Ligation: samples digested on the 15th of September

Ligation pairs: plasmid / insert

BBa_B0014 / BBa_I712669 ;
BBa_P1003/BBa_B0014 ;
BBa_J23100/BBa_B0014

First report:
Plasmid = 1µL
Insert = 6µL

Second report:
Plasmid = 3µL
Insert = 5µL

Third report:
Plasmid = 1µL
Insert = 5µL

NEB Enzymes

For each samples add sterilized water to obtain a maximum volume equals to 8µL.

Ligation mix (NEB):
9µL of T4 ligase + 9µL of T4 ligase buffer.
Add 2µL / tube.
Incubation 1h at RT.

Electroporation

Electroporation cuvettes = 2mm ; inoculums of electrocompetent E.coli DH5alpha= 40µL; pulse = 2,5KVolt ; 1h of incubation.

Spread 1mL of inoculums into a petri dish with LB + ampicillin (50mg/mL) (20/0,02mL).